anti kv 1 3 extracellular antibody  (Alomone Labs)

Journal: The Journal of comparative neurology

Article Title: Localization of Kv1.3 channels in presynaptic terminals of brainstem auditory neurons

doi: 10.1002/cne.22393

Figure Lengend Snippet: Gradients of Kv1.3 immunoreactivity in sagittal and coronal sections of the MNTB. A. Sagittal section of MNTB immunostained for Kv 1.3. (scale bar, 20 μm). B and C. Coronal (B) and sagittal (C) sections were divided into 5 equal zones for quantification

Article Snippet: Anti Kv 1.3 (extracellular) antibody was obtained from Alomone labs.

Techniques:

Journal: The Journal of comparative neurology

Article Title: Localization of Kv1.3 channels in presynaptic terminals of brainstem auditory neurons

doi: 10.1002/cne.22393

Figure Lengend Snippet: Electron microscopic immunogold labeling of Kv 1.3 in the MNTB of Wild-type and Kv 1.3 −/− mice. A and B. Examples of sections stained for Kv1.3 using 10 nm gold particles. Particles are localized primarily to pre-synaptic structures,

Article Snippet: Anti Kv 1.3 (extracellular) antibody was obtained from Alomone labs.

Techniques: Labeling, Mouse Assay, Staining

Journal: The Journal of comparative neurology

Article Title: Localization of Kv1.3 channels in presynaptic terminals of brainstem auditory neurons

doi: 10.1002/cne.22393

Figure Lengend Snippet: Co-localization of Kv 1.3 with syntaxin, synaptophysin and synaptotagmin. Double immunofluorescence labeling of Kv1.3 with syntaxin (top panels A – E ), synaptophysin ( center panels F – J ) and synaptotagmin ( bottom panels K – O ). Kv 1.3

Article Snippet: Anti Kv 1.3 (extracellular) antibody was obtained from Alomone labs.

Techniques: Immunofluorescence, Labeling

Journal: Annals of Clinical and Translational Neurology

Article Title: The potassium channel Kv1.3 as a therapeutic target for immunocytoprotection after reperfusion

doi: 10.1002/acn3.51456

Figure Lengend Snippet: Comparison of stroke severity in WT versus Kv1.3 −/− mice. (A) Infarct area and deficit score in 16‐week‐old WT ( n = 10) were compared to Kv1.3 −/− ( n = 14) male mice ( p

Article Snippet: The immunofluorescence staining in Figure was performed with a polyclonal guinea pig anti‐Kv1.3 antibody (1:100, AGP‐005 from Alomone Labs) and the same polyclonal rabbit Iba‐1 (1;1000) and CD3 (1:250) antibodies listed above; Secondary Abs: Alexa Fluor® 546‐goat anti‐guinea pig IgG (1:1000, ab150185 from Abcam) and Alexa Fluor®488‐goat anti‐rabbit IgG (1:1500, ab150077 from Abcam).

Techniques: Mouse Assay

Journal: Annals of Clinical and Translational Neurology

Article Title: The potassium channel Kv1.3 as a therapeutic target for immunocytoprotection after reperfusion

doi: 10.1002/acn3.51456

Figure Lengend Snippet: The Kv1.3 blocker PAP‐1 reduces infarction and improves neurological deficit in young and old WT female mice. (A) Infarct area and deficit score in vehicle ( n = 11) were compared to PAP‐1 (40 mg/kg; n = 15) treated 16‐week‐old female mice ( p = 0.007 for infarct, p = 0.016 for NES). (B) Infarct area and deficit score in vehicle ( n = 9) were compared to PAP‐1 (40 mg/kg; n = 9) treated 80‐week‐old female mice ( p = 0.032 for infarct, p = 0.003 for NES). Data are shown as whisker plots with data overlay. The boxes show mean ± SEM and the whiskers show confidence intervals.

Article Snippet: The immunofluorescence staining in Figure was performed with a polyclonal guinea pig anti‐Kv1.3 antibody (1:100, AGP‐005 from Alomone Labs) and the same polyclonal rabbit Iba‐1 (1;1000) and CD3 (1:250) antibodies listed above; Secondary Abs: Alexa Fluor® 546‐goat anti‐guinea pig IgG (1:1000, ab150185 from Abcam) and Alexa Fluor®488‐goat anti‐rabbit IgG (1:1500, ab150077 from Abcam).

Techniques: Mouse Assay, Whisker Assay

Journal: Annals of Clinical and Translational Neurology

Article Title: The potassium channel Kv1.3 as a therapeutic target for immunocytoprotection after reperfusion

doi: 10.1002/acn3.51456

Figure Lengend Snippet: Kv1.3 is expressed on microglia/macrophages and T cells in the infarct of an 80‐week‐old female mouse. (A) Sample immunofluorescence staining of 5‐ µ m thick paraffin sections from the 6‐mm slice of an 80‐week‐old female mouse on day 8 after tMCAO. (A) Kv1.3 staining colocalizes to Iba‐1 + cells. (B) Kv1.3 staining on smaller, CD3 + T cells (white arrowheads).

Article Snippet: The immunofluorescence staining in Figure was performed with a polyclonal guinea pig anti‐Kv1.3 antibody (1:100, AGP‐005 from Alomone Labs) and the same polyclonal rabbit Iba‐1 (1;1000) and CD3 (1:250) antibodies listed above; Secondary Abs: Alexa Fluor® 546‐goat anti‐guinea pig IgG (1:1000, ab150185 from Abcam) and Alexa Fluor®488‐goat anti‐rabbit IgG (1:1500, ab150077 from Abcam).

Techniques: Immunofluorescence, Staining

Journal: Annals of Clinical and Translational Neurology

Article Title: The potassium channel Kv1.3 as a therapeutic target for immunocytoprotection after reperfusion

doi: 10.1002/acn3.51456

Figure Lengend Snippet: PAP‐1 treatment and Kv1.3 deletion reduce Iba‐1 and CD3 staining in 80‐week‐old female mice. Serial paraffin sections (5 µ m thick) cut at 4 mm and the 6 mm from 80‐week‐old female mice were stained for Iba‐1 and CD3. (A) Percentage of Iba‐1‐positive pixels in the infarcted hemisphere. Vehicle‐treated WT ( n = 9), PAP‐1‐treated WT ( n = 9, p = 0.120 for 4‐mm section; p = 0.014 for 6‐mm section), and Kv1.3 −/− ( n = 18, p = 0.011 for 4‐mm section; p = 0.007 for 6‐mm section). (B) Percentage of CD3‐positive pixels in the infarcted hemisphere. Vehicle‐treated WT ( n = 9), PAP‐1‐treated WT ( n = 9, p = 0.049 for 4‐mm section; p = 0.027 for 6‐mm section), and Kv1.3 −/− ( n = 18, p

Article Snippet: The immunofluorescence staining in Figure was performed with a polyclonal guinea pig anti‐Kv1.3 antibody (1:100, AGP‐005 from Alomone Labs) and the same polyclonal rabbit Iba‐1 (1;1000) and CD3 (1:250) antibodies listed above; Secondary Abs: Alexa Fluor® 546‐goat anti‐guinea pig IgG (1:1000, ab150185 from Abcam) and Alexa Fluor®488‐goat anti‐rabbit IgG (1:1500, ab150077 from Abcam).

Techniques: Staining, Mouse Assay

Journal: Scientific Reports

Article Title: KCNE4-dependent functional consequences of Kv1.3-related leukocyte physiology

doi: 10.1038/s41598-021-94015-9

Figure Lengend Snippet: KCNE4 impaired Kv1.3 accumulation in the IS but did not disrupt IS formation. Human Jurkat T lymphocytes and human Raji B lymphocytes were used to generate cell conjugates. ( A – C ) Activated B-cells (10 µg/mL SEE toxin) were cocultured in the absence ( Ba – Bd ) or presence ( Ca – Cd ) of Jurkat cells, and confocal images were obtained. ( Aa – Ad ) Jurkat T-cells in the absence of B cells. Endogenous Kv1.3 (green), CD3 (marker of T-cells, red), and CD19 (marker of B-cells, blue) were detected. ( Ad , Bd , Cd ) merge panels. Note that triple colocalization (white) in Cd localizes Kv1.3 in the IS, as identified by CD3 staining ( Cb ). Bars are 20 µm. ( D ) Accumulation ratio of mGFP-Kv1.3 at the IS vs. KCNE4-mCherry total intensity (n = 40). ( E ) CD3 recruitment into the IS vs. KCNE4 intensity. The horizontal red line represents the threshold level (1.5) for Kv1.3 and CD3 accumulation in the IS. Values greater or less than 1.5 indicated positive or negative accumulation of proteins at the IS, respectively.

Article Snippet: Next, the cells were incubated with anti-KCNE4 (1/50, Proteintech) or anti-Kv1.3 extracellular (1/150, Alomone) antibodies with 10% goat serum in 0.05% Triton X-100 PBS-K+ for 1 h 45 min at RT.

Techniques: Marker, Staining

Journal: Scientific Reports

Article Title: KCNE4-dependent functional consequences of Kv1.3-related leukocyte physiology

doi: 10.1038/s41598-021-94015-9

Figure Lengend Snippet: KCNE4 expression impairs Kv1.3 surface expression and inhibits Kv currents in Jurkat T cells. Confocal imaging of Jurkat T cells transfected with YFP, Kv1.3YFP, KCNE4CFP and KCNE4CFP with YFP-Kv1.3. Nuclei were stained with DAPI (blue). ( Aa – Ad ) Jurkat nontransfected cells (control). ( Ba – Bd ) YFP-transfected cells (YFP). ( Ca – Cd ) Kv1.3YFP transfected cells. ( Da – Dd ) KCNE4CFP transfected cells. ( Ea – Ed ) Kv1.3YFP and KCNE4CFP cotransfected cells. ( Aa , Ba , Ca , Da , Ea ) Kv1.3 in green. ( Ab , Bb , Cb , Db , Eb ) KCNE4 in red. ( Ac , Bc , Cc , Dc , Ec ) DAPI in blue. Merged yellow indicates colocalization between green and red ( Ad , Bd , Cd , Dd , Ed ). Scale bar: 5 µm. ( F ) FRET analysis of the Kv1.3-KCNE4 protein interaction by flow cytometry in Jurkat T lymphocytes. Values are mean ± SE, n = 5–7, *p

Article Snippet: Next, the cells were incubated with anti-KCNE4 (1/50, Proteintech) or anti-Kv1.3 extracellular (1/150, Alomone) antibodies with 10% goat serum in 0.05% Triton X-100 PBS-K+ for 1 h 45 min at RT.

Techniques: Expressing, Imaging, Transfection, Staining, Flow Cytometry

Journal: Scientific Reports

Article Title: KCNE4-dependent functional consequences of Kv1.3-related leukocyte physiology

doi: 10.1038/s41598-021-94015-9

Figure Lengend Snippet: LPS-dependent activation of CY15 dendritic cells increases the abundance of Kv1.3 at the cell surface. CY15 cells were incubated in the presence (LPS) or the absence (control) of LPS for 24 h. Cells were first stained with WGA (membrane marker) and then immunolabeled against Kv1.3. ( A – D ) Control cells in the absence of LPS. ( E – H ) Cells treated with LPS. Green, Kv1.3; red, WGA; merged panels show colocalization between green and red. ( D , H ) Histogram of the pixel by pixel analysis of the section indicated by the arrow in ( C , G ), respectively. Bars represent 10 μm. ( I ) Mander's overlap coefficient (MOC) quantifying the degree of colocalization between Kv1.3 and membrane surface (WGA) staining. White bar, control; black bar, LPS. Values are mean ± SE of n > 30 cells. ***p

Article Snippet: Next, the cells were incubated with anti-KCNE4 (1/50, Proteintech) or anti-Kv1.3 extracellular (1/150, Alomone) antibodies with 10% goat serum in 0.05% Triton X-100 PBS-K+ for 1 h 45 min at RT.

Techniques: Activation Assay, Incubation, Staining, Whole Genome Amplification, Marker, Immunolabeling

Journal: Scientific Reports

Article Title: KCNE4-dependent functional consequences of Kv1.3-related leukocyte physiology

doi: 10.1038/s41598-021-94015-9

Figure Lengend Snippet: KCNE4 overexpression modulates Kv1.3-related physiological events in Jurkat T lymphocytes. Jurkat T cells were electroporated with KCNE4CFP, and positively transfected cells were selected for specific assays. ( A ) Kv1.3 and KCNE4 expression in Jurkat cells. ( B ) Percentage of Jurkat T cell proliferation. Cells were serum starved overnight and cultured for an additional 24 h in the presence of FBS. The alamarBlue dye was used. *p

Article Snippet: Next, the cells were incubated with anti-KCNE4 (1/50, Proteintech) or anti-Kv1.3 extracellular (1/150, Alomone) antibodies with 10% goat serum in 0.05% Triton X-100 PBS-K+ for 1 h 45 min at RT.

Techniques: Over Expression, Transfection, Expressing, Cell Culture

Journal: Scientific Reports

Article Title: KCNE4-dependent functional consequences of Kv1.3-related leukocyte physiology

doi: 10.1038/s41598-021-94015-9

Figure Lengend Snippet: Kv1.3 and KCNE4 are differentially expressed in leukocytes. The presence of Kv1.3 and KCNE4 expression was analyzed in human Jurkat T lymphocytes and mouse CY15 dendritic cells. ( A ) Kv1.3 and KCNE4 protein expression in leukocytes. HEK 293 cells were used as a negative control. Although Jurkat and CY15 dendritic cells shared Kv1.3 and KCNE4 expression, the abundance of KCNE4 in T cells was much lower and minimally detected by western blot. In addition, Kv1.5 was abundantly expressed in CY15 cells. Representative cropped blots, clearly separated by vertical white lines, are shown only for qualitative purposes. Voltage-dependent K + currents were elicited in Jurkat ( B ) and CY15 cells ( C ). Cells were held at -60 mV, and 250 ms pulse potentials were applied as indicated. ( D ) Representative confocal images of Kv1.3 ( Da and Dd , in green) and KCNE4 ( Db and De , in red) in Jurkat T lymphocytes ( Da – Dc ) and CY15 dendritic cells ( Dd – Df ). Scale bars: 10 µm. Given the limited expression of KCNE4 in T-cells, IPI was performed in Jurkat cells. ( E ) KCNE4 coimmunoprecipitated with Kv1.3 in dendritic cells. Lysates were immunoprecipitated against Kv1.3 (IP: Kv1.3) and immunoblotted (IB) against Kv1.3 and KCNE4. Upper panel: Kv1.3. Lower panel: KCNE4. SM: starting material. SN+: supernatant from the IP+. SN−: supernatant from the IP−. IP+: Immunoprecipitation in the presence of the anti-Kv1.3 antibody. IP−: Immunoprecipitated in the absence of the anti-Kv1.3 antibody. ( F ) Kv1.3 localized in lipid raft fractions from Jurkat T-cells. ( G ) Kv1.3 and KCNE4 did not localize in lipid rafts from CY15 dendritic cells. Lipid rafts were isolated, and low density (1) to high density (12) sucrose gradient fractions were analyzed by western blot. Flotilin indicated low-buoyancy lipid rafts, whereas clathrin identified nonfloating raft fractions.

Article Snippet: Next, the cells were incubated with anti-KCNE4 (1/50, Proteintech) or anti-Kv1.3 extracellular (1/150, Alomone) antibodies with 10% goat serum in 0.05% Triton X-100 PBS-K+ for 1 h 45 min at RT.

Techniques: Expressing, Negative Control, Western Blot, Immunoprecipitation, Isolation

Journal: Scientific Reports

Article Title: KCNE4-dependent functional consequences of Kv1.3-related leukocyte physiology

doi: 10.1038/s41598-021-94015-9

Figure Lengend Snippet: LPS-dependent activation increases the Kv1.3/KCNE4 ratio in CY15 dendritic cells. Cells were treated for 24 h with LPS (100 ng/ml), and the protein expression of selected K + channel proteins was studied at 0, 6 and 24 h. ( A ) Confocal images of CY15 cells stained with Kv1.3 upon LPS treatment. Scale bars: 20 µm. ( B ) Representative voltage-dependent K+ currents elicited in CY15 cells treated with (LPS 24 h) or without (LPS 0 h) LPS during 24 h. Cells were held at -60 mV and 250 ms pulses to +60 mV were applied. ( C ) Peak current densitiy (pA/pF) of K + currents from CY15 cells in the absence (0 h) or the presence (24 h) of LPS. White bars, LPS 0 h (control); black bars, LPS 24 h. Values are the mean ± SE of 4–6 cells. *p

Article Snippet: Next, the cells were incubated with anti-KCNE4 (1/50, Proteintech) or anti-Kv1.3 extracellular (1/150, Alomone) antibodies with 10% goat serum in 0.05% Triton X-100 PBS-K+ for 1 h 45 min at RT.

Techniques: Activation Assay, Expressing, Staining